PRODUCT INFORMATION
Pyrophosphatase,
Inorganic (from yeast)
Pub. No. MAN0011985
Rev. Date 21 September 2016 (Rev. B.00)
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Lot: _ Expiry Date: _
Store at -20 °C
Components
#EF0221
Pyrophosphatase, Inorganic
(from yeast)
10 U
0.1 U/µL
Storage (Dilution) Buffer
1 mL
www.thermofisher.com
For Research Use Only. Not for use in diagnostic procedures.
Description
Pyrophosphatase, Inorganic catalyzes the hydrolysis of
inorganic pyrophosphate to two orthophosphates. The
enzyme requires a divalent metal cation, with Mg2+
conferring the highest activity (1).
Applications
High yield RNA synthesis by in vitro transcription (2).
DNA polymerization reactions: preventing accumulation
of pyrophosphate (3, 4).
Removal of contaminant PPi in reagents used for SNP
genotyping by methods based on the detection of
pyrophosphate (5).
Source
E.coli cells with a cloned ppa gene of Saccharomyces
cerevisiae.
Molecular Weight
This enzyme is homodimer. It is consists of two identical
subunits of 32 kDa.
Definition of Activity Unit
One unit of the enzyme hydrolysis 1 µmol of inorganic
pyrophosphate in 1 min at 25 °C.
Enzyme activity is assayed in the following mixture:
100 mM Tris-HCl (pH 7.2), 2 mM MgCl2 and
2 mM inorganic pyrophosphate (PPi).
Storage (Dilution) Buffer
The enzyme is supplied in: 10 mM Tris-HCl (pH 7.5),
0.1 mM EDTA and 50% (v/v) glycerol.
Inhibition and Inactivation
Inhibitors: imidodiphosphate, α ω glycol disphosphates,
methanedial diphosphate, 1,2-ethanedial diphosphate (6).
Inactivation by heating is not complete, reliably removed
by spin column or phenol/chloroform extraction.
Note
The enzyme can be diluted with supplied storage
(dilution) buffer.
CERTIFICATE OF ANALYSIS
Endodeoxyribonuclease Assay
No detectable degradation was observed after
incubation of supercoiled plasmid DNA with
Pyrophosphatase, Inorganic.
Ribonuclease Assay
No detectable degradation was observed after incubation
of [3H]-RNA with Pyrophosphatase, Inorganic.
Labeled Oligonucleotide (LO) Assay
No detectable degradation after incubation of single
stranded and double stranded radiolabeled
oligonucleotides with Pyrophosphatase, Inorganic.
Quality authorized by: Jurgita Zilinskiene
(continued on back page)
References
1. Cooperman, B.S., The mechanism of action of yeast
inorganic pyrophosphatase, Meth. Enzymol., 87, 526-548,
1982.
2. Cunningham, P.R. and Ofengand, J., Use of inorganic
pyrophosphatase to improve the yield of in vitro transcription
reactions catalyzed by T7 RNA polymerase, Biotechniques, 9,
713-714, 1990.
3. Tabor, S., Richardson, C.C., DNA sequence analysis with a
modified bacteriophage T7 DNA polymerase. Effect of
pyrophosphorolysis and metal ions, J. Biol. Chem., 265,
8322-8328, 1990.
4. Dean, B.F., et al., Rapid amplification of plasmid and phage
DNA using phi29 DNA polymerase and multiply-primed
Rolling Circle amplification, Genome Res., 11, 1095-1099,
2001.
5. Zhou, G.H., et al., Quantitative detection of single nucleotide
polymorphisms for a pooled sample by a bioluminometric
assay coupled with modified primer extension reactions
(BAMPER), Nucleic Acids Res., 29, E93, 2001.
6. Sperow, J.W., et al., Yeast Inorganic Pyrophosphatase, The
Journal of Biological Chemistry, 6, 2062-2065, 1973.
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conveyed to the buyer to use the product or components of the product for purposes
including but not limited to provision of services to a third party, generation of
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research and development purposes, please contact [email protected] or
Out Licensing, Life Technologies Inc., 5791 Van Allen Way, Carlsbad, California
92008.
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This product is developed, designed and sold exclusively for research purposes and
in vitro use only. The product was not tested for use in diagnostics or for drug
development, nor is it suitable for administration to humans or animals.
Please refer to www.thermofisher.com for Material Safety Data Sheet of the product.
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